Background: Mashhad is a large pilgrimage city with over 20 million pilgrims and tourists annually. Some pilgrims donate their blood voluntarily during the pilgrimage. Objectives: The present study aimed to compare the prevalence of viral transfusion-transmitted infections (TTIs) between resident and pilgrim blood donors in Mashhad. Methods: We reviewed the records of all blood donors in Mashhad blood centers in 2011. The demographic data and the prevalence of TTIs, including HBV, HCV, HIV, and HTLV-1, were compared between 42821 donors from Mashhad and 15219 donors from other cities of Iran. The data were analyzed using Microsoft Excel 2010 and Epi-Info 6.0 software. Results:The mean age of residents and pilgrims was 34.6 ± 9.9 and 34.2 ± 10 years, respectively. The male to female ratio in the first group was higher than that in the second group (13.1 and 6.6, respectively). One-third of the residents and nearly two-thirds of the pilgrims were first-time blood donors. The most prevalent TTIs among the residents were HTLV-1 (0.39%) and HBV (0.33%); however, the frequencies of these infections were 0.24% and 0.57% among the pilgrims, respectively. The seroprevalence of HCV infection was 0.054% and 0.072% in the first and second groups, respectively. The HIV infection was observed only in one donor from Mashhad. Conclusions: A low prevalence of TTIs, particularly HCV and HIV infections, was found among blood donors from all parts of Iran. A higher prevalence of HTLV-1 infection among Mashhad donor population than among donors from other parts of Iran verified the virus endemicity in this region.